EVALUATION OF 3-ETHOXY-1,2,4-DITHIAZOLINE-5-ONE (EDITH) AS A NEW SULFURIZING REAGENT IN COMBINATION WITH LABILE EXOCYCLIC AMINO PROTECTING GROUPS FOR SOLID-PHASE OLIGONUCLEOTIDE SYNTHESIS

摘要

3-ethoxy-1,2,4-dithiazoline-5-one (EDITH) was recently introduced as an efficient sulfurizing reagent for solid-phase oligonucleotide synthesis, The successful syntheses were performed using standard base protecting groups (i.e. benzoyl for A and C, isobutyryl for G), which required deprotection in concentrated ammonium hydroxide at 55 degrees C for 15-18 h, We have explored the possibility of using EDITH in combination with fast deprotection chemistry (e.g. Expedite Chemistry using tert-butylphenoxy acetyl as a base protecting group). Surprisingly, poor synthesis performance was observed when syntheses were conducted with EDITH, Expedite Chemistry and standard synthesis cycle (i.e. Coupling-Thio-Cap), Potential G modification seemed to be the source of incompatibility since sequences containing no G or carrying isobutyryl-protected G residues could be synthesized with high efficiency. However, the deleterious G modification can be readily eliminated by inserting a capping step before the sulfurization reaction. Oligomers prepared with the Coupling-Cap-Thio-Cap cycle contained few phosphodiester contaminants as measured by P-31-NMR, anion-exchange HPLC and MALDI-TOF mass spectrometry, In addition to reducing deprotection time, this new combination also provides a mild method for the preparation of certain phosphorothioate oligomers that may be sensitive to prolonged ammonia treatment (e.g. thioated RNAs).